PLASMID ISOLATION AND ANALYSIS Part III Plasmid Isolation

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PLASMID ISOLATION AND ANALYSIS Part III Plasmid Isolation

PLASMID ISOLATION AND ANALYSIS Part III Plasmid Isolation

Plasmid Isolation Step 1 Place a QIAprep® spin column in the 2 ml collection

Plasmid Isolation Step 1 Place a QIAprep® spin column in the 2 ml collection tube. QIAprep® spin column collection tube PLASMID ISOLATION AND ANALYSIS: Part III

Plasmid Isolation Step 2 Use the small needle pipette to carefully remove all of

Plasmid Isolation Step 2 Use the small needle pipette to carefully remove all of the clear lysate/supernate from the “L” tube. PLASMID ISOLATION AND ANALYSIS: Part III

Plasmid Isolation Step 3 Add the clear lysate/supernate from tube “L” to the QIAprep®

Plasmid Isolation Step 3 Add the clear lysate/supernate from tube “L” to the QIAprep® column. PLASMID ISOLATION AND ANALYSIS: Part III

Plasmid Isolation Step 4 Centrifuge for 1 minute and discard the flow through. PLASMID

Plasmid Isolation Step 4 Centrifuge for 1 minute and discard the flow through. PLASMID ISOLATION AND ANALYSIS: Part III

Plasmid Isolation Step 5 Using a small needle pipette, add the contents of the

Plasmid Isolation Step 5 Using a small needle pipette, add the contents of the tube marked “Wash Buffer” (PE) to the QIAprep® spin column and centrifuge for 1 minute. PLASMID ISOLATION AND ANALYSIS: Part III

Plasmid Isolation Step 6 Discard the flow through, and centrifuge for an additional 1

Plasmid Isolation Step 6 Discard the flow through, and centrifuge for an additional 1 minute to remove residual wash buffer. PLASMID ISOLATION AND ANALYSIS: Part III

Plasmid Isolation Step 7 Transfer the QIAprep® spin column to the 1. 5 ml

Plasmid Isolation Step 7 Transfer the QIAprep® spin column to the 1. 5 ml tube marked “M” in Part II, step 6. QIAprep® spin column PLASMID ISOLATION AND ANALYSIS: Part III

Plasmid Isolation Step 8 Use a small needle pipette to elude the DNA by

Plasmid Isolation Step 8 Use a small needle pipette to elude the DNA by adding the contents of the tube marked sterile water (SW) to the QIAprep® spin column. PLASMID ISOLATION AND ANALYSIS: Part III

Plasmid Isolation Step 9 Let stand for 1 minute and centrifuge for 1 minute

Plasmid Isolation Step 9 Let stand for 1 minute and centrifuge for 1 minute to collect the DNA. PLASMID ISOLATION AND ANALYSIS: Part III

Plasmid Isolation Step 10 Label the “M” tube with your group’s name, seal the

Plasmid Isolation Step 10 Label the “M” tube with your group’s name, seal the tube with its original cap, and place in freezer (-20 o. C) until you are ready to analyze the extracted DNA in Part IV. PLASMID ISOLATION AND ANALYSIS: Part III